Phenylderivate as inhibitors of ATP citrate lyase

ABSTRACT

PCT No. PCT/EP93/01071 Sec. 371 Date Nov. 4, 1994 Sec. 102(e) Date Nov. 4, 1994 PCT Filed May 5, 1992 PCT Pub. No. WO93/22304 PCT Pub. Date Nov. 11, 1993.Compounds of the formula     &lt;IMAGE&gt;  (I)  where the substituents are defined in the specification useful in the treatment of hyperlipidemia.

This application is a 371 of PCT/EP93/01071 filed Apr. 29, 1993.

The present invention relates to certain novel compounds, processes andintermediates used in their preparation, pharmaceutical compositionscontaining them and their use in therapy.

It is now widely accepted that treatment of even moderate type IIhypercholesterolaemia results in a reduction in mortality and morbiditydue to coronary heart disease (CHD). Increased plasma concentrations oflow density lipoprotein (LDL), the hallmark of type IIhypercholesterolaemia are due to a variety of genetic and environmentalfactors resulting in increased LDL synthesis, decreased LDL catabolismor combinations of both. Current therapies for treatment ofhypercholesterolaemia are directed towards stimulation of LDL catabolism(bile acid sequestrants and HMGCoA reductase inhibitors) as well asinhibition of LDL synthesis (nicotinic acid and maxepa fish oil).

The present invention relates to a new class of compounds which areexpected to be of use in the treatment of hyperlipidaemia and preventingthe development of consequent disorders like atherosclerosis andpancreatitis, as well as treatment of metabolic disorders like obesity.The compounds act by inhibition of the enzyme ATP citrate lyase, soinhibiting cholesterol synthesis and fatty acid synthesis resulting inlowered plasma cholesterol and triglyceride levels. In particular, it isexpected that the compounds will be particularly useful in the treatmentof mixed hyperlipidaemia (type IIb).

The present invention therefore provides, in a first aspect, compoundsof structure (I) : ##STR2## in which, each group R¹ is independently alipophilic and/or electron withdrawing group;

n is 5 to 8; and

either R² and R³ are both hydrogen, R⁴ is hydrogen or hydroxy and R⁵ isCH(R⁶)R⁷ in which R⁶ is hydrogen or hydroxy and R⁷ is a carboxyl groupor a carboxylic acid ester group hydrolysable to a carboxyl group; or R⁴is hydrogen and R⁵ is hydrogen or hydroxy, R² is hydroxy and R³ is acarboxyl group or a carboxylic acid ester group hydrolysable to acarboxyl group; or R² and R³ are hydrogen and R⁴ and R⁵ together form agroup =C(R⁶)R⁷ in which R⁶ and R⁷ are as defined above,

and salts thereof.

The term "lipophilic and/or electron withdrawing group" refers to groupssuch as halogen, in particular chlorine, nitro, cyano, C₁₋₄ alkanoyl,optionally substituted phenylC₁₋₄ alkanoyl and fluorinated C₁₋₄ alkylsuch as trifluoromethyl. Other examples of such groups will be apparentto those skilled in the art. Suitable C₁₋₄ alkanoyl groups include CH₃CO-- and C₃ H₇ CO--. Suitable phenylC₁₋₄ alkanoyl groups include, forexample, phenylCO-(benzoyl).

"Carboxylic acid ester groups hydrolysable to a carboxyl group" asdefined for R³ and R⁷ include, for example, groups of formula C₂ R⁸ inwhich R⁸ is C₁₋₆ alkyl, benzyl, acetoxymethyl and pivaloyloxymethyl;preferably C₁₋₆ alkyl such as methyl. Other examples of such groups willbe apparent to those skilled in the art.

Suitably, each group R¹ is independently a lipophilic and/or electronwithdrawing group. Preferably each group R¹ is the same and positionedin the 2,3- or 2,4-positions of the ring, in particular the2,4-positions. More preferably each group R¹ is the same and is halogen,in particular chlorine in the 2,4-positions of the ring.

Suitably, n is 5 to 8, preferably 6 or 7.

Suitably, R² and R³ are both hydrogen, R⁴ is hydrogen or hydroxy and R⁵is CH(R⁶)R⁷ in which R⁶ is hydrogen or hydroxy and R⁷ is a carboxylgroup or a carboxylic acid ester group hydrolysable to a carboxyl group;or R⁴ is hydrogen and R⁵ is hydrogen or hydroxy, R² is hydroxy and R³ isa carboxyl group or a carboxylic acid ester group hydrolysable to acarboxyl group; or R² and R³ are hydrogen and R⁴ and R⁵ together form agroup =C(R⁶)R⁷ in which R⁶ and R⁷ are as defined above.

Preferably, R² and R³ are both hydrogen, R⁴ is hydroxy and R⁵ isCH(R⁶)R⁷ in which R⁶ is hydrogen and R⁷ is a carboxyl group.

Suitable salts of the compounds of structure (I) include, for example,basic salts, those formed by reaction with an appropriate base. Suchsalts include, for example, the sodium and potassium salts which can beprepared by methods well known to those skilled in the art, for example,the sodium salts can be formed by reaction with sodium hydroxide in anaqueous or non-aqueous medium.

The compounds of structure (I) can be prepared by procedures analogousto those known in the art. In a further aspect, there is thereforeprovided a process for preparing a compound of structure (I) whichcomprises:

(a) for compounds of structure (I) in which R² and R³ are both hydrogen,R⁴ is hydrogen or hydroxy and R⁵ is CH(R⁶)R⁷, lactonisation of acompound of structure (II): ##STR3## in which R¹, R⁷ and n are asdescribed for structure (I), and R⁹ is hydrogen or OR¹⁰ where R¹⁰ ishydrogen or C₁₋₄ alkyl, or

(b) for compounds of structure (I) in which R⁴ is hydrogen, R⁵ ishydrogen or hydroxy, R² is hydroxy and R³ is CO₂ H or a grouphydrolysable to CO₂ H, lactonisation of a compound of structure (III):##STR4## in which R¹, R⁷ and n are as described for structure (I), andR⁹ is hydrogen or OR¹⁰ where R¹⁰ is hydrogen or C₁₋₄ alkyl as definedabove, or

(c) for compounds in which R² and R³ are hydrogen, and R⁴ and R⁵together form a group ═CR⁶ R⁷ in which R⁶ and R⁷ are as described forstructure (I), lactonisation of a compound of structure (IV): ##STR5##in which R¹, R⁷ and n are as described for structure (I), and R⁹ ishydrogen or OR¹⁰ where R¹⁰ is hydrogen or C₁₋₄ alkyl, and optionallythereafter,

removing any protecting groups

forming a salt.

The lactonisation of a compound of structure (II) can be carried out ina suitable solvent in the presence of an acid catalyst at a temperatureof between ambient, and the boiling point of the solvent used, for aslong as is required for reaction to go to completion. For example, thereaction can be carried out in tetrahydrofuran, in the presence ofaqueous hydrochloride acid at a temperature of about 60° C. untilreaction is complete. Alternative solvent systems and suitable acidswill be apparent to those skilled in the art, for example the reactioncan be carried out in a non-aqueous solvent such as diethyl ether ortetrahydrofuran, in the presence of acid (for example sulphuric acid)impregnated silica gel as a catalyst.

The intermediate compounds of structure (II) can be prepared byreduction of compounds of structure (V): ##STR6## in which R¹, n, R⁷ andR⁹ are as described for structure (II). Suitable conditions for thereduction include hydrogenation over a suitable catalyst, in particularRaney nickel, in the presence of an acid such as hydroboric acid, in asuitable solvent such as a C₁₋₄ alkanol, in particular methanol,followed by reduction of the resulting intermediate ketone with, forexample, sodium borohydride in the presence of cerium chloride in asuitable solvent such as a C₁₋₄ alkanol, in particular methanol or,preferably, sodium acetoxyborohydride (prepared in situ from sodiumborohydride in acetic acid, or commercially available) in acetic acid assolvent.

The compounds of structure (V) can be prepared from compounds ofstructure (VI): ##STR7## in which R¹ and n are as described forstructure (II), by reaction with a compound of structure (VII): ##STR8##in which R⁷ is as described for structure (V). Suitable reactionconditions will be apparent to persons skilled in the art, for example,by reaction in a non-aqueous solvent, such as dichloromethane in thepresence of aqueous sodium hypochlorite and triethylamine.

The compounds of structure (VI) can themselves be prepared fromcommercially available starting materials as described in the specificexamples herein.

Alternatively, the compounds of structure (II) can be prepared byreduction of the compounds of structure (VIII): ##STR9## in which R¹, n,R⁷ and R⁹ are as described for structure (II). Suitable conditionsinclude, for example, hydrogenation over a suitable catalyst, inparticular Raney nickel, in the presence of an acid such as hydroboricacid, in a suitable solvent such as a C₁₋₄ alkanol, in particularmethanol, followed by hydrogenation over a noble metal catalyst such asplatinum oxide, and then reduction of the intermediate ketone so formedwith, for example, sodium borohydride in acetic acid.

Compounds of structure (VIII) can, themselves, be prepared by reactionof a compound of structure (IX): ##STR10## in which R¹ is as describedfor structure (I), with a compound of structure (X): ##STR11## in whichn, R⁷ and R⁹ are as described for structure (V).

Suitable reaction conditions will be apparent to those skilled in theart and include, for example, Wittig reaction conditions, using, forexample, a suitable base such as sodium hydride, in a suitable solventsuch as dimethyl sulphoxide, as hereinafter described. The compounds ofstructure (IX) can be prepared by standard procedures for thepreparation of Wittig reagents. It will be appreciated by those skilledin the art that in the preparation of compounds of structure (I) inwhich at least one of the groups R¹ is an alkanoyl or optionallysubstituted phenylalkanoyl group, a keto protected form of the structure(IX) is used, that is to say, a compound of structure (IXA): ##STR12##in which R is C₁₋₄ alkyl or phenyl and R¹ is as described for structure(I).

Alternatively, the compounds of structure (VIII) can be prepared byreaction of a compound of structure (XI): ##STR13## in which n, R⁷ andR⁹ are as described for structure (II), with a compound of structure(XII): ##STR14## in which R¹ is as described for structure (I) and X ishalogen, in particular iodine.

The reaction between a compound of structure (XI) and a compound ofstructure (XII) can be carried out under Heck conditions as hereinafterdescribed.

Compounds of structure (XI) can be prepared from compounds of structure(X) by reaction, for example, under Wittig conditions, using a suitablebase such as sodium hydride, in a suitable solvent such as dimethylsulphoxide as hereinafter described.

The compounds of structure (X) can be prepared from the compounds ofstructure (XIII): ##STR15## in which n, R⁷ and R⁹ are as described forstructure (VI), under Swern oxidation conditions as hereinafterdescribed.

Compounds of structure (XIII) can be prepared from the correspondingcompounds of structure (XIV): ##STR16## in which n is as described forstructure (I) by reaction with a compound of structure (VII) ashereinbefore described for the reaction between a compound of structure(VI) and a compound of structure (VII). Compounds of structure (XIV) canbe prepared by procedures known to those skilled in the art, for examplethe compound of structure (XIV) in which n is 6 can be prepared fromε-caprolactone (commercially available) as hereinafter described.

The lactonisation of a compound of structure (III) can be carried outunder standard conditions, for example as described above for thelactonisation of compounds of structure (II).

The intermediate compounds of structure (III) can themselves be preparedfrom compounds of structure (XV): ##STR17## in which R¹, n, R⁷ and R⁹are as described for structure (III), by reaction, for example, withosmium tetroxide, N-methylmorpholine N-oxide in aqueous acetone as asolvent.

Alternatively, compounds of structure (III) can be prepared byring-opening of an epoxide of formula (XVI): ##STR18## in which R¹, R²,n, R⁷ and R⁹ are as described for structure (III).

The compounds of structure (XVI) can themselves be prepared byepoxidation of compounds of structure (XV) under standard conditions.

The compounds of structure (XV) can be prepared from compounds ofstructure (XVII): ##STR19## in which R¹ and n are as described forstructure (XV) by, for example, oxidation under standard conditions,followed by reaction under Wittig conditions.

The compounds of structure (XVII) can be prepared from commerciallyavailable starting materials, using standard procedures as describedherein in the specific examples.

Lactonisation of compounds of structure (IV) can be carried out understandard conditions as hereinbefore described for the lactonisation ofthe compounds of structure (II) and (III).

Compounds of structure (IV) can be prepared from the correspondingcompounds of structure (XVIII): ##STR20## in which R¹, n, R⁷ and R⁹ areas described for structure (IV) by, for example, dehydration andhydrolysis using aqueous acid such as hydrochloric acid.

Compounds of structure (XVIII) can be prepared from the correspondingcompounds of structure (XIX): ##STR21## in which R¹, n, R⁷ and R⁹ are asdescribed for structure (XVIII) by reaction, for example, with hydrogencyanide.

Compounds of structure (XIX) can be prepared from compounds of structure(XX) and (XXI) under standard conditions as will be apparent to thoseskilled in the art. ##STR22## Compounds of structure (XX) and (XXI) arecommercially available or can be prepared by standard techniques.

The intermediate compounds of structures (II), (III), (IV), (V), (VI),(VIII), (IX), (IXA), (X), (XI), (XIII), (XIV), (XV), (XVI), (XVII),(XVIII), (XIX) and (XX) are themselves novel and form a further aspectof the invention.

It will be appreciated that the compounds of structure (I) contain oneor more asymmetric carbon atoms and are thus optically active compounds.As such, these compounds can exist as two (or more) optical isomers(enantiomers). Both the pure enantiomers, racemic mixtures (50% of eachenantiomer thereof) and unequal mixtures of the two are included withinthe scope of the present invention. Further, all diastereomeric formspossible (pure enantiomers and mixtures thereof) are within the scope ofthe invention.

Pure enantiomers of the individual compounds claimed herein are obtainedby resolution of mixtures using standard techniques, for example, viasalt formation using, for example,D-(-)-threo-2-amino-1-(4-nitrophenyl)propan-1,3-diol as hereinafterdescribed.

In addition, it will be apparent that the lactone form of structure (I)can also exist in the form of its open-chain equivalent of structure(IA) as follows: ##STR23## The present invention is intended to coverboth of these forms in which R¹, n, R², R³, R⁴ and R⁵ are as describedfor structure (I).

It will, of course, be appreciated that the lactone forms of structure(I) can be converted to the open chain forms (IA) under standardhydrolysis conditions.

Furthermore, it will be apparent that the compounds of structure (IA) inwhich R² and R³ are both hydrogen, R⁴ is hydrogen or hydroxy and R⁵ isCH(R⁶)R⁷ in which R⁶ is hydrogen or hydroxy and R⁷ is a carboxyl group,can also exist in their corresponding 6-membered lactone ring form. Such6-membered ring forms are also intended to be within the scope of thepresent invention.

Whilst the compounds of structure (I) are represented as lactonestructures, it is believed that in vivo, that is to say onadministration to subjects, the compounds convert to the open-chainform, and any carboxylic acid ester groups present convert to freecarboxyl groups, and it is this open chain hydrolysed form in which thecompounds bind to the ATP citrate lyase enzyme and exhibit the activityclaimed herein. The active form of the compounds of structure (I) istherefore believed to be the structure (IB) ##STR24## in which eachgroup R^(1b) is independently a lipophilic and/or electron withdrawinggroup;

n^(b) is 5 to 8; and

either R^(2b) and R^(3b) are both hydrogen, R^(4b) is hydrogen orhydroxy and R^(5b) is CH(R^(6b))CO₂ H in which R^(6b) is hydrogen orhydroxy; or R^(4b) is hydrogen and R^(5b) is hydrogen or hydroxy, R^(2b)is hydroxy and R^(3b) is CO₂ H ; or R^(2b) and R^(3b) are hydrogen andR^(4b) and R^(5b) together form a group ═C(R^(6b))CO₂ H, andpharmaceutically acceptable salts thereof.

The compounds of structure (IB) and the pharmaceutically acceptablesalts thereof have been found to be inhibitors of the enzyme ATP citratelyase and, as such, are expected to be of use in medicine in thetreatment of elevated serum cholesterol and triglyceride levels inmammals, including humans. In a still further aspect, the presentinvention therefore provides inhibitors of the enzyme ATP citrate lyasefor use in therapy, in particular for lowering serum triglyceride andcholesterol levels in the treatment of mixed hyperlipidaemia (type(II)b). More particularly, the present invention provides compounds ofstructure (IB) and their pharmaceutically acceptable salts, for use intherapy, in particular for lowering serum triglyceride and cholesterollevels in the treatment of mixed hyperlipidaemia (Type (II)b). Inaddition, the compounds, that is to say, inhibitors of ATP citratelyase, in particular the compounds of structure (IB), are expected toexhibit a beneficial effect in preventing the development of consequentdisorders like atherosclerosis and pancreatitis as well as the treatmentof metabolic disorders like obesity.

In therapeutic use, the compounds of the present invention are usuallyadministered in a standard pharmaceutical composition. The presentinvention therefore provides, in a further aspect, pharmaceuticalcompositions comprising a compound of structure (I) or apharmaceutically acceptable salt thereof and a pharmaceuticallyacceptable carrier.

The compounds of structure (I) and their pharmaceutically acceptablesalts which are active when given orally can be formulated as liquids,for example, syrups, suspensions or emulsions, tablets, capsules andlozenges.

A liquid formulation will generally consist of a suspension or solutionof the compound or pharmaceutically acceptable salt in a suitable liquidcarrier(s), for example, ethanol, glycerine, non-aqueous solvent, forexample, polyethylene glycol, oils, or water with a suspending agent,preservative, flavouring or colouring agent.

A composition in the form of a tablet can be prepared using any suitablepharmaceutical carrier(s) routinely used for preparing solidformulations. Examples of such carriers include magnesium stearate,starch, lactose, sucrose and cellulose.

A composition in the form of a capsule can be prepared using routineencapsulation procedures. For example, pellets containing the activeingredient can be prepared using standard carrier and then filled into ahard gelatin capsule; alternatively, a dispersion or suspension can beprepared using any suitable pharmaceutical carrier(s), for example,aqueous gums, celluloses, silicates or oils and the dispersion orsuspension then filled into a soft gelatin capsule.

Typical parenteral compositions consist of a solution or suspension ofthe compound or pharmaceutically acceptable salt in a sterile aqueouscarrier or parenterally acceptable oil, for example, polyethyleneglycol, polyvinyl pyrrolidone, lecithin, arachis oil or sesame oil.Alternatively, the solution can be lyophilised and then reconstitutedwith a suitable solvent just prior to administration.

A typical suppository formulation comprises a compound of formula (I),or a pharmaceutically acceptable salt thereof, which is active whenadministered in this way, with a binding and/or lubricating agent suchas polymeric glycols, gelatins or cocoa butter or other low meltingvegetable or synthetic waxes or fats.

Preferably, the composition is in unit dose form such as a tablet orcapsule.

Each dosage unit for oral administration contains preferably from 1 to250 mg (and for parenteral administration contains preferably from 0.1to 25 mg) of a compound of the formula (I) or a pharmaceuticallyacceptable salt thereof calculated as the free base.

The present invention also provides a method of lowering serumtriglyceride and cholesterol levels which comprises administering to amammal in need thereof an effective amount of an inhibitor of the enzymeATP citrate lyase; and a method of lowering serum triglyceride andcholesterol levels which comprises administering to a subject in needthereof, an effective amount of a compound of structure (I) or (IB), ora pharmaceutically acceptable salt thereof.

The daily dosage regimen for an adult patient may be, for example, anoral dose of between 1 mg and 1000 mg, preferably between 0.1 mg and 250mg, or an intravenous, subcutaneous, or intramuscular dose of between0.1 mg and 100 mg, preferably between 0.1 mg and 25 mg, of the compoundof formula (I) or a pharmaceutically acceptable salt thereof calculatedas the free base, the compound being administered 1 to 4 times per day.Suitably, the compounds will be administered for a period of continuoustherapy, for example, for a week or more.

In addition, the compounds of the present invention can beco-administered (together or sequentially) with further activeingredients, for example and other hypercholesterolaemic agents such asbile acid sequestrants, ACAT inhibitors and other drugs for thetreatment of cardiovascular disease.

The following examples illustrate the invention. Temperatures arerecorded in degrees centigrade.

EXAMPLE 1 ±(3R*, 5S*)3-Carboxy-11-(2,4-dichlorophenyl)-3,5-dihydroxyundecanoic acid, disodiumsalt

(a) Methyl 7-(2,4-Dichlorophenyl)-6-heptenoate

Sodium hydride (60% dispersion in oil, 14.8 g, 369.4 mmol) was washedwith petroleum ether 40°-60° C., then heated to 80° C. in dimethylsulphoxide (180 ml) under argon until gas evolution ceased. The solutionwas cooled to 0° C. in an ice-bath, and a solution of5-carboxypentyltriphenylphosphonium bromide (82.4 g, 180.2 mmol) indimethylsulphoxide (380 ml) was added. The solution was stirred for 0.5h at room temperature, then cooled to 0° C. A solution of2,4-dichlorobenzaldehyde (31.5 g, 180.2 mmol) in dimethylsulphoxide (80ml) was added, and the mixture stirred at room temperature for 1 h, thenpoured into aqueous HCl. This mixture was extracted with ether. Allextracts were washed with water, saturated aqueous NaCl and dried overMgSO₄. The solvent was removed under vacuum.

Concentrated H₂ SO₄ (2 ml) was added to a solution of the residue inmethanol (300 ml), and this was stirred at room temperature for 15 h.Saturated aqueous NaHCO₃ was added until pH neutral and solvent wasremoved under vacuum. The residue was partitioned between aqueous NaHCO₃and ether. The ether layer was washed with aqueous HCl, water, saturatedaqueous NaCl, and dried (MgSO₄). The solvent was removed under vacuum,the residue extracted with 10% ether/petroleum ether 40°-60° C., and theextracts filtered. The solvent was removed under vacuum, and the residuepurified by chromatography on silica gel (10-30% ether/petroleum ether40°-60° C.) to give the title compound (40.2 g) as an oil, comprising amixture of E and Z isomers, which was used without further purification.

(b) 7-(2,4-Dichlorophenyl)-6-hepten-1-ol

Di-isobutylaluminium hydride (1.0M in dichloromethane, 308 ml, 308 mmol)was injected into a stirred solution of methyl7-(2,4-dichlorophenyl)-6-heptenoate (40.2 g, 140 mmol) indichloromethane (200 ml) at -78° C. under argon. After 5 min, thesolution was warmed to 0° C., stirred for 0.5 h, then cooled again to-78° C. Water (102 ml) was injected slowly, while allowing the mixtureto warm to room temperature. When solid had precipitated, ethyl acetate(400 ml) was added, then excess NaHCO₃, and the mixture stirredvigorously for 0.25 h. The solids were filtered off, the solvent removedunder vacuum, and the residue purified by chromatography on silica gel(30-70% ether/petroleum ether 40°-60° C.) to give the title compound(31.7 g, 85%) as an oil, comprising a mixture of E and Z isomers.

(c) 7-(2,4-Dichlorophenyl)-1-heptanol

A solution of 7-(2,4-dichlorophenyl)-6-hepten-1-ol (31.7 g, 122 mmol) inmethanol (150 ml) was shaken under hydrogen (50 psi) with platinum oxide(1.65 g, added in portions) until no starting material could be detectedby NMR spectroscopy. The catalyst was filtered off, and the solventremoved under vacuum. The residue was dissolved in ether, and thesolution filtered through a pad of silica gel. The solvent was removedunder vacuum to give the title compound (30.5 g, 96%) as an oil.

(d) 7-(2,4-Dichlorophenyl) heptanaldoxime

Dimethylsulphoxide (15.2 ml, 214 mmol) was added slowly to a stirredsolution of oxalyl chloride (9.35 ml, 107 mmol) in dichloromethane (150ml) at -78° C. under argon. After 5 min, a solution of7-(2,4-dichlorophenyl)-1-heptanol (20.0 g, 76.6 mmol) in dichloromethane(100 ml) was added by cannula. After stirring 0.5 h at -78° C.,triethylamine (47 ml, 337 mmol) was injected. The mixture was stirred 5min, allowed to warm to room temperature, then poured into 1M aqueousNaHSO₄. The product was extracted with ether. The extracts were washedwith water, saturated aqueous NaCl, then the solvent removed undervacuum.

A solution of the crude aldehyde in ether (120 ml) was added to astirred suspension of hydroxylamine hydrochloride (17.0 g, 245 mmol) inwater (10 ml) at 0° C., followed by aqueous Na₂ CO₃ (2.7M, 50 ml, 135mmol). The mixture was stirred at room temperature for 2.5 h, pouredinto water, and extracted with ether. The extracts were washed withwater, saturated aqueous NaCl, and dried (MgSO₄). The solvent wasremoved under vacuum, and the residue purified by chromatography onsilica gel (30-50% ether/petroleum ether 40°-60° C.) to give the titlecompound (17.3 g, 82%) as a mixture of E and Z isomers.

(e) ±5-(Carbomethoxymethyl)-3-[6-(2,4-dichlorophenyl)-hexyl]-5-methoxycarbonyl-4,5-dihydroisoxazole

Aqueous NaOCl (2.0M, 340 ml, 680 mmol) and triethylamine (2.5 ml, 18.0mmol) were added in 4 portions separately over 40 h to a stirredsolution of 7-(2,4-dichlorophenyl)-heptanaldoxime (17.3 g, 63.1 mmol),and dimethyl itaconate (23.0 g, 145 mmol) in dichloromethane (200 ml).The mixture was stirred vigorously at room temperature over this period,then poured into water and extracted with ether. The extracts werewashed with water, saturated aqueous NaCl, and dried (MgSO₄). Thesolvent was removed under vacuum, and the residue purified bychromatography on silica gel (30-60% ether/petroleum ether 40°-60° C.)to give the title compound (19.5 g, 72%) as an oil.

(f) ±Methyl11-(2,4-dichlorophenyl)-3-hydroxy-3-methoxycarbonyl-5-oxo-undecanoate

A solution of±5-(carbomethoxymethyl)-3-[6-(2,4-dichlorophenyl)hexyl]-5-methoxycarbonyl-4,5-dihydroisoxazole(19.5 g, 45.3 mmol) and boric acid (8.39 g, 136 mmol) in methanol wasshaken with Raney nickel (50% slurry in water, 8 g) under hydrogen (50psi) at room temperature for 2 h. The catalyst was removed byfiltration, and most of the solvent removed under vacuum. The mixturewas diluted with water, and extracted with ethyl acetate. The extractswere washed with water, saturated aqueous NaCl, and dried (MgSO₄) . Thesolvent was removed under vacuum, and the residue purified bychromatography on silica gel (50-80% ether/petroleum ether 40°-60° C.)to give the title compound (16.7 g, 85%) as an oil.

(g) ±(3R*,5S*) 3-Carboxy-11-(2,4-dichlorophenyl)-3,5-dihydroxyundecanoicacid, disodium salt

Sodium borohydride (1.47 g, 38.8) was added in portions to a stirredsolution of ± methyl11-(2,4-dichlorophenyl)-3-hydroxy-3-methoxycarbonyl-5-oxo-undecanoate(16.0 g, 36.9 mmol) and cerium (III) chloride heptahydrate (14.4 g, 38.8mmol) in methanol (200 ml) at 0° C. The solution was stirred for 0.5 h,then quenched with aqueous HCl. The mixture was diluted with water, andextracted with ether. The extracts were washed with water, saturatedaqueous NaCl, dried (MgSO₄), and the solvent removed under vacuum.Aqueous NaOH (1.5M, 100 ml, 150 mmol) was added to a stirred solution ofthe crude hydroxy ester in ethanol (100 ml) at 0° C. The mixture wasstirred for 4 h, diluted with ethanol (200 ml), and filtered. The solidwas recrystallised (aqueous ethanol) to give the title compound (11.9 g,a monohydrate, 69%) as a white solid, m.p. indeterminate.

C₁₈ H₂₂ Cl₂ O₆ Na₂.H₂ O Found C 46.03%, H 5.20% Requires C 46.07%, H5.15%.

EXAMPLE 2 ±(3R*, 5S*) 3-Carboxymethyl-5-[6-(2,4-dichlorophenyl)hexyl]3-hydroxytetrahydrofuran-2-one

A mixture of ±(3R*, 5S*)3-carboxy-11-(2,4-dichlorophenyl)-3,5-dihydroxyundecanoic acid, disodiumsalt (11.8 g, 25.1 mmol), aqueous HCl (3M, 200 ml), and tetrahydrofuran(200 ml) was heated at 60° C. for 6 h, then cooled. Most of thetetrahydrofuran was removed under vacuum. The residual mixture wasdiluted with water, and extracted with ether. The extracts were washedwith water, saturated aqueous NaCl, and dried (MgSO₄). The solvent wasremoved under vacuum, and the residue recrystallised (ether/petroleumether 40°-60° C.) to give the title compound (8.80 g, 90%) as a whitesolid, m.p. 77°-79° C.

C₁₈ H₂₂ Cl₂ O₅ Found C 55.34%, H 5.64% Requires C 55.54%, H 5.70%.

EXAMPLE 3 ±(E)-3-Carboxy-11-(2,4-dichlorophenyl)-5-hydroxy-2-undecenoicacid

(a) ±Methyl 11-(2,4-dichlorophenyl)-5-hydroxy-3-oxo-undecanoate

Dimethylsulphoxide (0.381 ml, 5.36 mmol) was injected dropwise into astirred solution of oxalyl chloride (0.234 ml, 2.68 mmol) indichloromethane (4 ml) at -78° C. under argon. After 5 min, a solutionof 7-(2,4-dichlorophenyl)-1-heptanol (see example 1, 500 mg, 1.91 mmol)in dichloromethane (4 ml) was added by cannula. The mixture was stirredfor 0.5 h, then triethylamine (1.17 ml, 8.40 mmol) added. The reactionwas allowed to warm to room temperature, then poured into aqueousNaHSO₄. The mixture was extracted with ether, and the extracts washedwith water, saturated aqueous NaCl, and dried (MgSO₄). The solvent wasremoved under vacuum.

Methyl acetoacetate (0.247 ml, 2.29 mmol) was added dropwise to astirred suspension of sodium hydride (61 mg, 2.52 mmol) intetrahydrofuran (2 ml) at 0° C. under argon. The solution was stirred0.5 h at room temperature, cooled to 0° C., and n-butyllithium (2.5M inhexanes, 1.01 ml, 2.52 mmol) injected. This solution was stirred 0.25 hat room temperature, cooled to -78° C., and a solution of the crudealdehyde in tetrahydrofuran (3 ml) added. The mixture was stirred for0.3 h at -78° C., allowed to warm to room temperature, then poured intoaqueous NaHSO₄, and extracted with ether. The extracts were washed withwater, saturated aqueous NaCl, and dried (MgSO₄). The solvent wasremoved under vacuum, and the residue purified by chromatography onsilica gel (60-100% ether/petroleum ether 40°-60° C.) to give the titlecompound (563 mg, 79%) as an oil.

(b) ±(E)-3-Carboxy-11-(2,4-dichlorophenyl)-5-hydroxy-2-undecenoic acid

Aqueous KH₂ PO₄ (1.12M, 13 ml, 14.6 mmol) was added to a stirred mixtureof ± methyl 11-(2,4-dichlorophenyl)-5-hydroxy-3-oxo-undecanoate (547 mg,1.46 mmol), KCN (951 mg, 14.6 mmol), and ether (8 ml). The mixture wasstirred vigorously for 18 h, then conc. aqueous HCl (1.25 ml, 14.6 mmol)added dropwise. The ether layer was removed, and the aqueous washed withether. All extracts were concentrated under vacuum.

The crude cyanohydrin was heated under reflux in aqueous HCl (7.7M) for3.5 h. The reaction was cooled, diluted with water, and the mixtureextracted with ether. The extracts were concentrated under vacuum.Aqueous NaOH (1M, 4 ml ) was added to a stirred solution of the residuein ethanol (5 ml) at 0° C. After 1 h, the precipitate was filtered off,and the filtrate diluted with water. This solution was washed withether, acidified to pH 1.5, and extracted with ether. The extracts werewashed with water, saturated aqueous NaCl, and dried (MgSO₄). Thesolvent was removed under vacuum, and the residue recrystallised(ether/petroleum ether 40°-60° C.) to give the title compound (110 mg,19%) as a white solid, m.p. 92°-93° C.

C₁₈ H₂₂ Cl₂ O₅ Found C 55.66%, H 5.72% Requires C 55.54%, H 5.70%.

EXAMPLE 4 ±(3R*,5R*)3-Carboxy-11-(2,4-dichlorophenyl)-3,5-dihydroxyundecanoic acid, disodiumsalt

The precipitated sodium salts obtained during the preparation of thecompound of example 3b were recrystallised (aqueous ethanol) to give a1:1 mixture of the title compound and its diastereoisomer (131 mg, 19%)as a white solid, m.p. indeterminate.

C₁₈ H₂₂ Cl₂ O₆.Na₂,1.3H2O Found C 45.54%, H 5.07% Requires C 45.54%, H5.23%.

EXAMPLE 5 ±(4R*,5S*)4-Carboxy-5-[8-(2,4-dichlorophenyl)octyl]-4-hydroxytetrahydrofuran-2-one

(a) (E)-Methyl 12-(2,4-Dichlorophenyl)-3-methoxycarbonyl-3-dodecenoate

Dimethylsulphoxide (1.37 ml, 19.3 mmol) was added dropwise to a stirredsolution of oxalyl chloride (0.844 ml, 9.67 mmol) in dichloromethane (35ml) at -78° C. under argon. After 5 min, a solution of9-(2,4-dichlorophenyl)-1-nonanol (2.00 g, 6.91 mmol: prepared inanalogous fashion to 7-(2,4-dichlorophenyl)-1-heptanol described inexample 1) in dichloromethane (10 ml ) was added by cannula. After afurther 0.5 h, triethylamine (4.24 ml, 30.4 mmol) was injected, and thereaction allowed to warm to room temperature. The mixture was pouredinto aqueous NaHSO₄ and products extracted with ether. The extracts werewashed with water, saturated aqueous NaCl, and dried (MgSO₄). Thesolvent was removed under vacuum. A solution of the crude aldehyde and1,2di(methoxycarbonyl)ethylenetriphenylphosphorane (4.21 g, 10.4 mmol)in toluene (25 ml) was heated at 100° C. for 24 h, then cooled. Thesolvent was removed under vacuum, and the residue purified bychromatography on silica gel (10-40% ether/petroleum ether 40°-60° C.)to give the title compound (2.46 g, 86%) as an oil.

(b) ±(3R*,4S*) Methyl12-(2,4-Dichlorophenyl)-3,4-dihydroxy-3-methoxycarbonyldodecanoate

A mixture of (E) methyl12-(2,4-dichlorophenyl)-3-methoxycarbonyl-3-dodecenoate (1.50 g, 3.61mmol), osmium tetroxide (0.229 ml of a 2% solution in t-butanol, 0.018mmol), N-methylmorpholine-N-oxide (634 mg, 5.42 mmol), water (2 ml), andacetone (2 ml) was stirred at room temperature for 64 h. Aqueous NaHSO₃(1.1M, 6 ml) was added, and the mixture filtered, after 20 min, througha plug of silica gel. The silica gel plug was washed with ether, thenthe filtrate washed with aqueous HCl, water, and saturated aqueous NaCl.After drying (MgSO₄), the solvent was removed under vacuum, and theresidue purified by chromatography on silica gel (50-100%ether/petroleum ether 40°-60° C.) to give the title compound (1.33 g),contaminated with the 5-ring lactone.

(c) ±(4R*,5S*) 4-Carboxy-5-[8-(2,4-dichlorophenyl)octyl]-4-hydroxytetrahydrofuran-2-one

±(3R*,4S*) Methyl12-(2,4-dichlorophenyl)-3,4-dihydroxy-3-methoxycarbonyl-3-dodecanoate(0.95 g, 2.11 mmol) was heated under reflux in aqueous HCl (7.7M) for3.5 h. The mixture was cooled, diluted with water, and extracted withether. The extracts were washed with aqueous NaOH, then the aqueousextracts washed with ether, acidified (aqueous HCl), and extracted againwith ether. The extracts were washed with water, saturated aqueous NaCl,and dried (MgSO₄). The solvent was removed under vacuum. The crudediacid was stirred in ether with silica gel (5 g) impregnated with 0.5ml 2M aqueous H₂ SO₄ at room temperature for 18 h, then the mixturefiltered, and the solvent removed under vacuum from the filtrate. Theresidue was recrystallised (dichloromethane/petroleum ether 40°-60° C.)to give the title compound (485 mg, 57%) as a gummy solid.

C₁₉ H₂₄ Cl₂ O₅ Found C 56.55%, H 6.04% Requires C 56.59%, H 6.00%.

EXAMPLE 6 ±(3R*,4S*) 3-Carboxy-11-(2 4-dichlorophenyl)-34-dihydroxyundecanoic acid

±(3R*,4S*) Methyl11-(2,4-dichlorophenyl)-3,4-dihydroxy-3-methoxycarbonylundecanoate (189mg, 0.434 mmol: prepared in analogous fashion to the higher homologuedescribed in example 5) was heated under reflux in aqueous HCl (7.7M)for 3 h. The solution was cooled, diluted with water, and extracted withether. The extracts were washed with aqueous NaOH, then the aqueousphase washed with ether, and acidified (aqueous HCl). The mixture wasextracted with ether again. The extracts were washed with water,saturated aqueous NaCl, and dried (MgSO₄). The solvent was removed undervacuum, and the residue recrystallised (ether/petroleum ether 40°-60°C.) to give the title compound (115 mg, 68%) as a white solid, m.p.104°-105° C.

C₁₈ H₂₄ Cl₂ O₆ Found C 53.00%, H 5.93% Requires C 53.08%, H 5.94%.

EXAMPLE 7 ±(4R*,5R*)4-Carboxy-5-[8-(2,4-dichlorophenyl)octyl]-4-hydroxytetrahydrofuran-2-one

a) ±(3R*,4S*) Methyl12-(2,4-Dichlorophenyl)-3,4-epoxy-3-methoxycarbonyldodecanoate

(E)-Methyl 12-(2,4-dichlorophenyl)-3-methoxycarbonyl-3-dodecenoate (686mg, 1.65 mmol, described in example 4) and 3-chloroperbenzoic acid (5.7g of 55% grade, 18.2 mmol) were heated under reflux in dichloromethane(15 ml) for 24 h. The mixture was cooled and poured into aqueous NaHCO₃/Na₂ SO₃. This mixture was stirred for 10 min, then extracted withdichloromethane. The extracts were washed with aqueous NaHC₃, water,saturated aqueous NaCl, and dried (MgSO₄). The solvent was removed undervacuum, and the residue purified by chromatography on silica gel (20-40%ether/petroleum ether 40-60%) to give the title compound (369 mg, 52%)as an oil.

(b) ±(4R*,5R*,)5-[8-(2,4-Dichlorophenyl)octyl]-4-hydroxy-4-methoxycarbonyltetrahydrofuran-2-one

±(3R*,4S*) Methyl12-(2,4-dichlorophenyl)-3,4-epoxy-3-methoxycarbonyldodecanoate (314 mg,0.728 mmol) was heated at reflux in aqueous H₂ SO₄ (7.5M) for 3.5 h. Thesolution was diluted with water, and extracted with ether. The extractswere washed with water, saturated aqueous NaCl, and dried (MgSO₄). Thesolvent was removed under vacuum. Diazomethane, generated from diazald(450 mg, 2.10 mmol) and aqueous KOH (10.7M), was passed in a stream ofether saturated nitrogen through a solution of the crude acid in 10%methanol/ether (10 ml) until a yellow colouration was seen in thereaction flask. Excess diazomethane was quenched with acetic acid, thenthe solvent removed under vacuum. The residue was purified bychromatography on silica gel (50-80% ether/petroleum ether 40°-60° C.)to give the title compound (198 mg, 65%) as an oil.

(c) ±(4R*,5R*) 4-Carboxy-5-[8-(2,4-dichlorophenyl)octyl]-4-hydroxytetrahydrofuran-2-one

Aqueous NaOH (1M, 4 ml, 4.0 mmol) was added dropwise to a stirredsolution of ±(4R*,5R*,) 5-[8-(2,4-dichlorophenyl)-octyl]-4-hydroxy-4-methoxycarbonyltetrahydrofuran-2-one (198 mg, 0.474 mmol) in methanol(4 ml) at 0° C. The solution was stirred at room temperature for 24 h,diluted with water, and washed with ether. The aqueous phase wasacidified (aqueous HCl), and extracted with ether. The extracts werewashed with water, saturated aqueous NaCl, and dried (MgSO₄). Thesolvent was removed under vacuum. The crude diacid was stirred withsilica gel (3 g), impregnated with 2M aqueous H₂ SO₄ (0.15 ml), in etherfor 1 h. The mixture was filtered, and the solvent removed from thefiltrate under vacuum to give the title compound (107 mg), (δ 200 MHz,CDCl₃) 7.34-7.10 (3H, m), 4.66 (1H, dd), 3.25 (1H, d), 2.77 (1H, d),2.67 (2H, t), 1.95-1.20 (14H, m), contaminated with a small amount of±(E) (4R*,5R*)4-carboxy-5-[8-(2,4-dichlorophenyl)-7-octenyl]-4-hydroxytetrahydrofuran-2-one.

EXAMPLE 8 ±(2R*,3R*,5S*)3-Carboxy-11-(2,4-dichlorophenyl)-2,3,5-trihydroxyundecanoic acid,disodium salt

(a) ±(3R*,5S*)3-Carbomethoxymethyl-5-[6-(2,4dichlorophenyl)hexyl]-3-hydroxytetrahydrofuran-2-one

Diazomethane, generated from diazald (1.37 g, 6.38 mmol) and 60% aqueousKOH (6 ml) in carbitol (6 ml) and ether (6 ml), was bubbled in a streamof ether saturated nitrogen through a solution of ±(3R*,5S*)3-carboxymethyl-5-[6-(2,4-dichloro-phenyl)hexyl]-3-hydroxytetrahydrofuran-2-one(1.24 g, 3.19 mmol, see example 2) in 10% methanol/ether (12 ml). When ayellow colour appeared in the solution, the excess diazomethane wasquenched with acetic acid, then the solvent removed under vacuum. Theresidue was purified by chromatography on silica gel (50-100%ether/petroleum ether 40°-60° C.) to give the title compound (1.15 g,90%) as an oil.

(b) ±(1'R*,3R*,5S*)3-[(Carbomethoxy)hydroxymethyl]-5-[6(2,4-dichlorophenyl)hexyl]-3-hydroxytetrahydrofuran-2-one

n-Butyllithium (3.27 ml, 8.18 mmol, 2.5M in hexanes) was injected into astirred solution of hexamethyldisilazane (1.73 ml, 8.18 mmol) intetrahydrofuran (15 ml) at 0° C. under argon. After 5 min, the solutionwas cooled to -78° C. and a solution of ±(3R*, 5S*)3-(carbomethoxymethyl)-5-[6-(2,4dichlorophenyl)hexyl]-3-hydroxytetrahydrofuran-2-one (1.15 g, 2.85mmol) in tetrahydrofuran (10 ml) was added by cannula slowly. After 1 hat -78° C., a solution of 2-(benzenesulphonyl)-3-phenyloxaziridine (1.07g, 4.09 mmol) in tetrahydrofuran (7 ml) was added by cannula. Thesolution was stirred at -78° C. for 2 h, at -50° C. for 2.5 h, thenallowed to warm to 0° C.

Aqueous HCl was added, and the mixture extracted with ether. Theextracts were washed with water, saturated aqueous NaCl, and dried(MgSO₄). The solvent was removed under vacuum, and the residue purifiedby chromatography on silica gel (60-100% ether/petroleum ether 40°-60°C.) to give the title compound (236 mg, 20%) as an oil, as well as the(1'R*,3S*,5R*)-diastereoisomer (145mg, 12%), also as an oil.

(C) ±(2R*,3R*,5S*)3-Carboxy-11-(2,4-dichlorophenyl)-2,3,5-trihydroxyundecanoic acid,disodium salt

Aqueous NaOH (1M, 2.25 ml, 2.25 mmol) was added dropwise to a stirredsolution of ±(1'R*,3R*,5S*)3-[(carbomethoxy)-hydroxymethyl]-5-[6-(2,4-dichlorophenyl)hexyl]-3-hydroxytetrahydrofuran-2-one(235 mg, 0.56 mmol) in methanol (6 ml) at 0° C. The solution was stirred10 min at 0° C., then 3 h at room temperature.

Methanol was removed under vacuum, the residue diluted with water, andacidified with aqueous HCl. The mixture was extracted with ether, andthe extracts washed with water. The solvent was removed under vacuum,and the wet residue dissolved in ethanol (20 ml). Aqueous NaOH (1M, 1.4ml, 1.4 mmol) was added. The mixture was allowed to stand for 45min,then boiled and cooled to 0° C. The solid was filtered off, washed with5% water/ethanol and dried to give the title compound (181 mg, 69%),m.p. indeterminate, contaminated with 4% of the (2R*,3S*,5R*)diastereoisomer.

C₁₈ H₂₂ Cl₂ Na₂ O₇. 0.84H₂ O Found C 44.76%, H 4.55%. Requires C 44.82%,H 4.95%.

EXAMPLE 9 ±(2R*,3S*,5R*)3-Carboxy-11-(2,4-dichlorophenyl)-2,3,5-trihydroxyundecanoic acid,disodium salt

Aqueous NaOH (1M, 1.38 ml, 1.38 mmol) was added dropwise to a stirredsolution of ±(1'R*,3S*,5R*)3-[(carbomethoxy)-hydroxy-methyl]-5-[6-(2,4-dichlorophenyl)hexyl]-3-hydroxytetrahydrofuran-2-one(145 mg, 0.346 mmol) in methanol (4 ml) at 0° C. The solution wasstirred at 0° C. for 10 min, then at room temperature for 3 h. Methanolwas removed under vacuum, and the aqueous residue acidified with aqueousHCl. The mixture was extracted with ether, and the extracts washed withwater. The solvent was removed under vacuum. The residue was dissolvedin ethanol (15 ml), and the solution filtered. Aqueous NaOH (1M, 0.8 ml,0.8 mmol) was added, the mixture allowed to stand for 45 min, thenboiled and cooled to 0° C. The solid was filtered off, washed with 5%water/ethanol, and dried to give the title compound (111 mg, 69%), m.p.indeterminate, contaminated with 20% of the (2R*,3R*,5S*)diastereoisomer.

C₁₈ H₂₂ Cl₂ Na₂ O₇.0.9H₂ O Found C 44.66%, H 4.57%. Requires C 44.72%, H4.96%.

EXAMPLE 10 ±(3R*,5R*)3-(Carboxymethyl)-5-[6-(2,4-dichlorophenyl)hexyl]tetrahydrofuran-2-oneand its diastereoisomer

(a) 8-(2,4-Dichlorophenyl)-1-octene

Dimethylsulphoxide (0.761 ml, 10.72 mmol) was added dropwise to astirred solution of oxalyl chloride (0,468 ml, 5.36 mmol) indichloromethane (10 ml) at -78° C. under argon. After 3 min, a solutionof 7-(2,4-dichlorophenyl)-1-heptanol (1.00 g, 3.83 mmol, see example 1c)in dichloromethane (5 ml) was added by cannula. After a further 30 min,triethylamine (2.35 ml, 16.9 mmol) was injected and the mixture warmedto room temperature, poured into aqueous HCl, and extracted with ether.The extracts were washed with water, saturated aqueous NaCl, and dried(MgSO₄). The solvent was removed under vacuum.

n-Butyllithium (2.5M, 3.37 ml, 8.43 mmol) was added to a stirredsuspension of methyltriphenylphosphonium bromide (2.74 g, 7.66 mmol) intetrahydrofuran (15 ml) at 0° C. under argon. The solution was stirredfor 0.5 h, then cooled to -78° C. A solution of the crude aldehyde intetrahydrofuran (5 ml) was added by cannula. After 5 min, the solutionwas warmed to room temperature, stirred for 1 h, then poured intoaqueous HCl and extracted with ether. The extracts were washed withwater, saturated aqueous NaCl, and dried (MgSO₄). The solvent wasremoved under vacuum, and the residue triturated with 10%ether/petroleum ether 40°-60° C. The extracts were filtered through aplug of silica gel, and the filtrate concentrated under vacuum to givethe title compound (697 mg), sufficiently pure to use in the nextreaction.

(b) ±2-[6-(2,4-Dichlorophenyl)hexyl]oxirane

m-Chloroperbenzoic acid (50% grade, 1.38 g, 4.01 mmol) was added inportions to a vigorously stirred mixture of8-(2,4-dichlorophenyl)-1-octene (687 mg, 2.67 mmol), saturated aqueousNaHCO₃ (15 ml), and dichloromethane (10 ml) at 0° C. The mixture wasstirred for 5 min at 0° C., at room temperature for 1 h, then pouredinto water, and extracted with ether. The extracts were washed withaqueous Na₂ SO₃ /NaHCO₃, water, saturated aqueous NaCl, and dried(MgSO₄). The solvent was removed under vacuum and the residue purifiedby chromatography on silica gel (5-20% ether/petroleum ether 40°-60° C.)to give the title compound (265 mg, 25%, 3 steps) as an oil.

(c) ±(3R*,5R*)3-(Carbomethoxymethyl)-5-[6-(2,4-dichlorophenyl)hexyl]tetrahydrofuran-2-oneand its diastereoisomer.

n-Butyllithium (2.5M, 1.22 ml, 3.06 mmol) was added to a stirredsolution of hexamethyldisilazane (0.646 ml, 3.06 mmol) intetrahydrofuran (5 ml) at 0° C. under argon. The solution was stirred 5min, then cooled to -78° C. A solution of diethyl succinate (0.462 ml,2.78 mmol) in tetrahydrofuran (3 ml) was added slowly by cannula, andthe mixture stirred for 30 min. A solution of±2-[6-(2,4-dichlorophenyl)hexyl]oxiirane (253 mg, 0.926 mmol) intetrahydrofuran (3 ml) was then added by cannula, followed immediatelyby boron trifluoride etherate (0.125 ml, 1.02 mmol). The mixture wasallowed to warm to room temperature slowly, then poured into aqueous HCland extracted with ether. The extracts were washed with water, saturatedaqueous NaCl, and dried (Na₂ SO₄). The solvent was removed under vacuumand the residue heated at reflux in 25% aqueous HCl for 4 h. Aftercooling and diluting with water, the mixture was extracted with ether.The extracts were washed with water, saturated aqueous NaCl, and dried(MgSO₄). The solvent was removed under vacuum and the residue purifiedby chromatography on silica gel (ether, then 0.5% acetic acid/ether) togive the crude acid lactone.

Diazomethane, generated from diazaid (227 mg, 1.06 mmol), 60% KOH (2ml), carbitol (2 ml) and ether (2 ml), was bubbled in an ether saturatedstream of nitrogen through a solution of the crude acid in 10%methanol/ether (5 ml) until excess diazomethane was observed, thenacetic acid added to quench. The solvent was removed under vacuum andthe residue purified by chromatography on silica gel (50-70%ether/petroleum ether 40°-60° C.) to give the title compound (185 mg,52%) as an oil, comprising a 1:1 mixture of diastereoisomers.

(d) ±(3R*,5R*)3-(Carboxymethyl)-5-[6-(2,4dichlorophenyl)hexyl]tetrahydrofuran-2-oneand its diastereoisomer

Aqueous NaOH (1M, 1.43 ml, 1.43 mmol) was added slowly to a stirredsolution of the diastereoisomers of±3-(carbomethoxymethyl)-5-[6-(2,4-dichlorophenyl)hexyl]tetrahydrofuran-2-one(185 mg, 0. 478 mmol) in methanol (5 ml) at 0° C. After 5 min at 0° C.,the mixture was stirred at room temperature for 6 h, then poured intoaqueous HCl and extracted with ether. The solvent was removed undervacuum and the residue heated at 60° C. in a 1:1 mixture of 3M aqueousHCl and tetrahydrofuran for 6 h. The mixture was cooled and extractedwith ether. The extracts were washed with water, saturated aqueous NaCl,and dried (MgSO₄). The solvent was removed under vacuum to give an oil,which slowly crystallised. The solid was recrystallised (ether/petroleumether 40°-60° C.) to give the title compound (126 mg, 71%), m.p. 57°-59°C.

C₁₈ H₂₂ Cl₂ O₄ Found C 57.73%, H 5.81%. Requires C 57.92%, H 5.94%.

EXAMPLE 11 ±(1'R*,3S*,5R*)3-[Carboxy(hydroxy)methyl]-5-[6-(2,4-dichlorophenyl)hexyl]-3-hydroxytetrahydrofuran-2 -one

Employing the method of example 2, substituting ±(2R*,3S*,5R*)3-carboxy-11-(2,4-dichlorophenyl)-2,3,5-trihydroxyundecanoic acid,disodium salt for ±(3R*,5S*)3-carboxy-11-(2,4-dichlorophenyl)-3,5-dihydroxyundecanoic acid, disodiumsalt, gave the title compound.

C₁₈ H₂₂ Cl₂ O₆ Found C 53.16% H 5.35% Requires C 53.35% H 5.47%.

EXAMPLE 12 ±(3R*,5S*)5-{6-[2,4-Bis(trifluoromethyl)phenyl]hexyl}-3-carboxymethyl-3-hydroxytetrahydrofuran-2-one

(a) 2,4-Bis(trifluoromethyl)benzyltriphenylphosphonium Bromide

N-Bromosuccinimide (3.17 g, 17.8 mmol) was added in portions to astirred solution of 2,4-bis(trifluoromethyl)benzyl alcohol (3.96 g, 16.2mmol) and triphenylphosphine (4.68 g, 7.8 mmol) in dichloromethane (40ml) at 0° under argon, and the solution stirred at room temperature for90 h. The solvent was removed under reduced pressure, and the residueloaded on to a pad of silica gel. The product was eluted with 50%ether/petroleum ether 40°-60° C.

A solution of the crude bromide and triphenylphosphine (4.25 g, 16.2mmol) in toluene (35 ml) was heated under reflux for 3 h, then cooled.The solid was filtered off, washed with ether, and dried to give thetitle compound (7.23 g, 78%) as a solid, m.p. 239°-244° C.

(b)±5-(Carbomethoxymethyl)-3-(5-hydroxypentyl)-5-methoxycarbonyl-4,5-dihydroisoxazole

A solution of di-isobutylaluminium hydride in dichloromethane (1.0M, 125ml, 125 mmol) was injected over 15 min into a stirred solution ofε-caprolactone (13.0 g, 114 mmol) in dichloromethane (100 ml) at -78°under argon. The solution was stirred for 20 min, then the cold bathremoved, and water (41 ml, 2.28 mol) injected. The mixture was stirredvigorously while allowing to warm to room temperature. When the solidhad separated, ethyl acetate was added, followed by excess sodiumbicarbonate, and stirring was continued for 10 min. The solids werefiltered off through a pad of hyflo, and the solvent removed from thefiltrate under reduced pressure.

Aqueous Na₂ CO₃ (2M, 100 ml, 200 mmol) was added slowly with vigorousstirring to a mixture of the crude hydroxyaldehyde and hydroxylaminehydrochloride (25.3 g, 364 mmol) in ether (200 ml). The mixture wasstirred for 3 h after the addition, then NaCl added to saturate theaqueous layer. The ether layer was separated, and the solvent removedunder reduced pressure. The aqueous layer was extracted with isobutanol.

The organic extracts were combined with the residue from the initialextract, washed with saturated aqueous NaCl, and dried (MgSO₄). Thesolvent was removed under reduced pressure, and traces of isobutanolremoved by azeotropic distillation with toluene.

Aqueous sodium hypochlorite (˜15%, 175 ml, ˜350 mmol) was added dropwiseto a stirred solution of the crude oximes, dimethyl itaconate (22.1 g,140 mmol) and triethylamine (1 ml, 7.17 mmol) in dichloromethane (100ml) cooled in a water bath. The mixture was stirred for 1 h, thenfiltered through hyflo. The organic layer was separated, and the aqueousextracted with dichloromethane. The extracts were washed with water,saturated aqueous NaCl, and dried (MgSO₄), then the solvent removedunder reduced pressure. Column chromatography of the residual oil onsilica gel (50-100% ether/petroleum ether 40°-60° C., then ethylacetate) gave the title compound (23.1 g, 69%) as an oil.

(c)±3-{6-[2,4-Bis(trifluoromethyl)phenyl]-5-hexenyl}-5-(carbomethoxymethyl)-5-methoxycarbonyl-4,5-dihydroisoxazole

Dimethylsulphoxide (0,692 ml, 9.74 mmol) was injected dropwise into astirred solution of oxalyl chloride (0,425 ml, 4.87 mmol) indichloromethane (10 ml) at -78° under argon. After 2 min, a solution of±5-(carbomethoxymethyl)-3-(5-hydroxypentyl)-5-methoxycarbonyl-4,5-dihydroisoxazole (1.00 g, 3.48mmol) in dichloromethane (5 ml) was added by cannula, and the mixturestirred for 30 min. Triethylamine (2.13 ml, 15.3 mmol) was injected,then the mixture allowed to warm to room temperature, poured intoaqueous HCl, and extracted with dichloromethane. The extracts werewashed with water, saturated aqueous NaCl, and dried (MgSO₄). Thesolvent was removed under reduced pressure, and the crude aldehyde driedby evaporation of a toluene solution. Sodium hydride (60% oilsuspension, 146 mg, 3.65 mmol) was washed with petroleum ether 40°-60°under argon, then heated with dimethylsulphoxide (5 ml) at 70-90° untilall solid had dissolved. After cooling in a water bath, a solution of2,4-bis(trifluoromethyl)benzyltriphenylphosphonium bromide (1.98 g, 3.48mmol) in dimethylsulphoxide (15 ml) was added by cannula, and themixture stirred at room temperature for 15 min. A solution of the crudealdehyde in dimethylsulphoxide (5 ml) was added, the mixture stirred for20 h, then poured into aqueous HCl and extracted with ether. Theextracts were washed with water, saturated aqueous NaCl, and dried(MgSO₄). The solvent was removed under reduced pressure, and the residuepurified by chromatography on silica gel (50-70% ether/petroleum ether40°-60° ) to give the title compound (0.56 g, 33%) as a mixture of E andZ isomers.

(d) ±Methyl11-[2,4-Bis(trifluoromethyl)phenyl]-3-hydroxy3-methoxycarbonyl-5-oxo-undecanoate.

A solution of±3-{6-[2,4-bis(trifluoromethyl)phenyl]-5-hexenyl}-5-(carbomethoxymethyl)-5-methoxycarbonyl-4,5-dihydroisoxazole(659 mg, 1.33 mmol) and boric acid (247 mg, 3.99 mmol) in methanol/water(10:1, 10 ml) was shaken with Raney nickel (˜300 mg) under hydrogen at40 psi for 4 h. The hydrogen was replaced with nitrogen, then thecatalyst filtered off through hyflo. The filtrate was diluted withwater, and extracted with ethyl acetate. The extracts were washed withwater, saturated aqueous NaCl, and dried (MgSO₄). The solvent wasremoved under reduced pressure.

A solution of the crude ketone in methanol (10 ml) was shaken withplatinum oxide (36 mg, 0.159 mmol) under hydrogen at 50 psi for 8 h,then the hydrogen replaced with nitrogen and the catalyst filtered offthrough hyflo. The solvent was removed under reduced pressure and theresidue columned on silica gel (50-80% ether/petroleum ether 40°-60° )to give the title compound (543 mg, 82%) as an oil.

(e) ±(3R*,5S*)5-{6-[2,4-Bis(trifluoromethyl)phenyl]-hexyl}-3-carboxymethyl-3-hydroxytetrahydrofuran-2-one

Following the procedures described in examples 1 (g) and 2, substituting±methyl 11-[2,4-Bis(trifluoromethyl)phenyl]-3-hydroxy-3-methoxycarbonyl-5-oxo-undecanoatefor ± methyl 11(2,4-dichlorophenyl)-3-hydroxy-3-methoxycarbonyl-5-oxo-undecanoate, gave the title compoundas a solid, m.p. 70°-73° C.

C₂₀ H₂₂ F₆ O₅ Found C 52.50% H 4.82% Requires C 52.64% H 4.86%.

EXAMPLE 13 ±(3R*,5S*) 3-Carboxy-11-(4-chloro-2-trifluoromethylphenyl)-3,5-dihydroxyundecanoic acid, disodium salt

(a)±5-Carbomethoxymethyl-3-hex-5-enyl-5-methoxycarbonyl4,5-dihydroisoxazole

The Swern oxidation of±5-(carbomethoxymethyl)-3-(5-hydroxypentyl)-5-methoxycarbonyl-4,5-dihydroisoxazole(1.00 g, 3.48 mmol) was carried out as described in example 12(c).

A solution of n-butyllithium in hexane (2.5M, 1.75 ml, 4.38 mmol) wasadded dropwise to a stirred suspension of methyltriphenylphosphoniumbromide (1.4 9 g, 4.18 mmol) in tetrahydrofuran (19 ml) at 0° underargon. After 30 min, the solution was cooled to -78° C., and a solutionof the crude aldehyde in tetrahydrofuran (6 ml) was added by cannula.The mixture was stirred 5 min at -78° C., 30 min at 0° C., then pouredinto aqueous HCl and extracted with ether. The extracts were washed withwater, saturated aqueous NaCl, and dried (MgSO₄). The solvent wasremoved under reduced pressure, and the residue purified by columnchromatography on silica gel (40-60% ether/petroleum ether 40°-60° ) togive the title compound (570 mg, 58%) as an oil.

(b)±5-Carbomethoxymethyl-3-[6-(4-chloro-2-trifluoromethylphenyl)-5-hexenyl]-5-methoxycarbonyl-4,5-dihydroisoxazole

A solution of 5-chloro-2-iodobenzotrifluoride (613 mg, 2.00 mmol),±5-carbomethoxymethyl-3-hex-5-enyl-5-methoxycarbonyl-4,5-dihydroisoxazole(567 mg, 2.00 mmol) and tributylamine (0.477 ml, 2.00 mmol) inN-methylpyrrolidinone (4 ml) was heated with palladium acetate (5 mg,0.022 mmol) under argon in an oil bath at 110° C. for 18 h, then cooled,poured into aqueous HCl, and extracted with ether. The extracts werewashed with water, saturated aqueous NaCl, and dried (MgSO₄). Thesolvent was removed under reduced pressure, and the residue purified bycolumn chromatography on silica gel (50-80% ether/petroleum ether40°-60° ) to give the title compound (534 mg) contaminated with otherisomeric olefins.

(c) ±(3R*,5S*)3-Carboxy-11-(4-chloro-2-trifluoromethyl-phenyl)-3,5-dihydroxyundecanoicacid, disodium salt

Following the procedures described in example 12(d) and (e),substituting±5-carbomethoxymethyl-3-[6-(4-chloro-2-trifluoromethylphenyl)-5-hexenyl]-5-methoxycarbonyl-4,5dihydroisoxazolefor±3-{6-[2,4-bis(trifluoromethyl)phenyl]-5-hexenyl)-5-(carbomethoxymethyl)-5-methoxycarbonyl4,5-dihydroisoxazole,gave the title compound as a solid, m.p. >250° C.

C₁₉ H₂₂ ClF₃ Na₂ O₆.0.5H₂ O Found C 46.02% H 4.77% Requires C 46.21% H4.69%.

EXAMPLE 14 ±(3R*,5S*)11-(2-Acetyl-4-chlorophenyl)-3-carboxy-3,5-dihydroxyundecanoic acid,disodium salt

(a) 5-Chloro-1-(1,1-ethylenedioxyethyl)-2-methylbenzene

Acetyl chloride (8.43 ml, 118.5 mmol) was injected into a stirredmixture of aluminium trichloride (15.8 g, 118.5 mmol) anddichloromethane (50 ml) under argon. When the solid had dissolved,4-chlorotoluene (4.67 ml, 39.5 mmol) was injected. The solution wasstirred for 20 h, then poured on to ice. The mixture was partitionedbetween salted water and ether, and the organic extracts washed withwater, saturated aqueous NaCl, and dried (MgSO₄). The solvent wasremoved under reduced pressure, and the residue purified by columnchromatography on silica gel (5-15% ether/petroleum ether 40°-60° ) togive a mixture (˜57:43) of 2-acetyl and 3-acetyl-4-chlorotoluene.

A solution of the isomeric ketones, ethanediol (16.15 ml, 289 mmol), andp-toluenesulphonic acid monohydrate (550 mg, 2.89 mmol) in toluene (50ml) was heated at reflux for 2 h, using a Dean and Stark separator toremove water. The solution was cooled, poured into water, and extractedwith ether. The extracts were washed with water, saturated aqueous NaCl,and dried (MgSO₄). The solvent was removed under reduced pressure, andthe residue purified by column chromatography on silica gel (2-8%ether/petroleum ether 40°-60° ) to give the title compound (2.98 g, 35%)as an oil.

(b) 4-Chloro-2-(1,1-ethylenedioxyethyl)benzyltriphenylphosphoniumBromide

A solution of 5-chloro-l-(1,1-ethylenedioxyethyl)-2-methylbenzene (2.97g, 14.0 mmol) and N-bromosuccinimide (2.74 g, 15.4 mmol) in carbontetrachloride (30 ml) was heated at reflux for 3 h, then cooled. Thesolvent was removed under reduced pressure, and the residue trituratedwith 20% ether/petroleum ether 40°-60° C. The extracts were filteredthrough a pad of silica gel, and the solvent removed from the filtrateunder reduced pressure. The residue was purified by columnchromatography on silica gel (3-9% ether/petroleum ether 40°-60° C.).

A solution of the bromide and triphenylphosphine (3.84 g, 14.7 mmol) intoluene (30 ml) was heated at reflux for 3 h, then cooled. The solid wasfiltered off, washed with ether, and dried under reduced pressure togive the title compound (5.94 g, 77%), m.p. 204°-209° C.

(c) ±Methyl11-[4-Chloro-2-(1,1-ethylenedioxy)phenyl]-3-hydroxy-3-methoxycarbonyl-5-oxo-undecanote

Following the procedures described in example 12(c) and (d),substituting4-chloro-2-(1,1-ethylenedioxyethyl)benzyltriphenylphosphonium bromidefor 2,4-bis(trifluoromethyl)benzyltriphenylphosphonium bromide, gave thetitle compound as an oil.

(d) ±(3R*,5S*)11-(2-Acetyl-4-chlorophenyl)-3-carboxy-3,5-dihydroxyundecanoic acid,disodium salt

Sodium borohydride (155 mg, 4.10 mmol) was added slowly in portions tostirred acetic acid (6 ml) cooled in a cold water bath. The solution wasstirred for 5 min, then a solution of ± methyl11-[4-chloro-2-(1,1-ethylenedioxy)phenyl]-3-hydroxy-3-methoxycarbonyl-5-oxo-undecanoate(510 mg, 1.05 mmol) in acetic acid (3 ml) was added. The mixture wasstirred at room temperature for 1 h, then poured into salted water andextracted with ether. The extracts were washed with water, saturatedaqueous NaCl, and dried (MgSO₄). The solvent was removed under reducedpressure.

A solution of the crude reduced product in trifluoroacetic acid/water(10:1, 10 ml) was stirred at room temperature for 1.5 h, then dilutedwith water and extracted with ether. The extracts were washed withwater, saturated aqueous NaCl, and dried (MgSO₄). The solvent and excesstrifluoroacetic acid were removed under reduced pressure.

Aqueous NaOH (1M, 3.15 ml, 3.15 mmol) was added dropwise to a stirredsolution of the crude ketone in ethanol (20 ml) at 0° C. The mixture wasallowed to warm to room temperature, then stirred for 18 h. Ethanol (20ml) was added, and the solid filtered off. Recrystallisation fromaqueous ethanol gave the title compound (337 mg, 70%) as a solid,m.p. >250° C.

C₂₀ H₂₅ ClNa₂ O₇ Found C 52.01% H 5.65% Requires C 52.35% H 5.49%.

EXAMPLES 15 and 16

Following the procedures described in example 14, substituting theappropriate acid chloride for acetyl chloride, gave the followingcompounds:

EXAMPLE 15 ±(3R*,5S*)11-(2-Benzoyl-4-chlorophenyl)-3-carboxy-3,5-dihydroxyundecanoic acid,disodium salt (0.3H₂ O)

C₂₅ H₂₇ ClO₇ Na₂.0.32H₂ O Found C 57.01% H 5.29% Requires C 57.01% H5.29%.

EXAMPLE 16 ±(3R*,5S*)11-(2-Butanoyl-4-chlorophenyl)-3-carboxy-3,5-dihydroxyundecanoic acid,disodium salt

C₂₂ H₂₉ ClNa₂ O₇.0.33H₂ O Found C 53.61% H 6.10% Requires C 53.61% H6.07%.

EXAMPLES 17-20 Resolution of Compounds of Examples 1 and 2 EXAMPLE 17(+) (3R*,5S*)3-Carboxymethyl-5-[6-(2,4-dichlorophenyl)-hexyl]-3-hydroxytetrahydrofuran-2-one

The lactone of example 2 (3.4 g, 8.9 mmol) was dissolved inethanol-water (96:4) (20 ml). To this was added a solution ofD-(-)-threo-2-amino-1-(4-nitrophenyl)propan-1, 3-diol (1.89 g, 8.9 mmol)in the same solvent (70 ml). After 3 hr, the crystallised solid (1.42 g)was collected and dried in vacuo. This salt was suspended in water and2M HCl added to pH 1-2. This was extracted with ether (3×), the combinedextracts washed with water (1×) and dried over MgSO₄. Concentration gavea white solid (0.9 g) which was recrystallised from CHC1₃ /hexane togive the pure (+)-enantiomer of example 2 (0.57 g); mp 80°-80.5° C.;[α]D²⁵ =+19.4 (c=0.5% w/v; EtOH).

C₁₈ H₂₂ Cl₂ O₅ Found C 55.54% H 5.61% Requires C 55.54% H 5.70%

EXAMPLE 18 (+) (3R*,5S*)3-Carboxy-11-(2,4-dichlorophenyl)-3,5-dihydroxyundecanoic acid, disodiumsalt

The compound of example 17 (160 mg) was dissolved in ethanol (2.5 ml)and 5% aqueous NaOH solution (0.64 ml) in water (1.86 ml) added withstirring. After ca 10 min, the precipitated solid was collected, washedwith 1:1EtOH-H₂ O and recrystallised from water-ethanol to give the(+)-enantiomer of example 1 (100 mg), [α]D²⁵ =+23.3 (c=0.16% w/v; H₂ O).

C₁₈ H₂₂ Cl₂ O₆ Na₂.H₂ O Found C 45.84% H 4.91% Requires C 46.07% H 5.15%

EXAMPLE 19 (-) (3R*,5S*)3-Carboxymethyl-5-[6-(2,4-dichlorophenyl)-hexyl]-3-hydroxytetrahydrofuran-2-one

By an analogous procedure to example 17, employing the enantiomericamine, (L-(+)-threo-2-amino-1-(4-nitrophenyl)propan-1,3-diol), gave thepure (-)-enantiomer of example 2, [α]D²⁵ =-19.5 (c=0.5% w/v; EtOH).

C₁₈ H₂₂ Cl₂ O₅ Found C 55.37% H 5.58% Requires C 55.54% H 5.70%

EXAMPLE 20

(-) (3R*,5S*) 3-Carboxy-11-(2 4-dichlorophenyl)-3 5-dihydroxyundecanoicacid, disodium salt

Substituting the compound of example 19 (100 mg) in the procedure ofexample 18 gave the (-)-enantiomer of example 1 (65 mg), [α]D²⁵ =-20.6(c=0.1% w/v; H₂ O).

C₁₈ H₂₂ Cl₂ O₆ Na₂.H₂ O Found C 45.69% H 4.98% Requires C 46.07% H 5.15%

Data:

1. RAT ATP CITRATE LYASE (ACL) ASSAY

Purification of ATP Citrate lyase-human and rat enzymes

(i) rat enzyme

Male Wistar rats were fasted for 24 h, then fed on a high carbohydratediet for 72 h prior to removal of the livers. ATP Citrate lyase wasprepared according to the method of Wraight et al (Anal. Biochem., 1985,144, 604-609) with modifications for large scale purification accordingto Wells (Eur. J. Biochem., 1991, 199, 163-168). Protein obtained bythis method was pure as judged by SDS-PAGE.

(ii) human enzyme

Human ATP citrate lyase was prepared as described in European Journal ofBiochemistry, 1992, 204, 491-99, with modifications for large scalepurification according to Wells as referred to above. Protein obtainedby this method was pure as judged by SDS-PAGE.

Assay of ATP Citrate lyase in the presence of inhibitors

ATP Citrate lyase activity was assayed at 25° C. by reducing theoxaloacetate produced with malate dehydrogenase and NADH whilemonitoring at 340 nm using a Beckman DU50 spectrophotometer (accordingto the method of Linnet al (J. Biol. Chem., 1979, 254, 1691-1698)).Briefly, ATP citrate lyase (human or rat) was added to a 1 ml cuvettecontaining 50 mM Tris/HCl, pH=8.0, 0.2 mM NADH, 10 mM MgCl₂, 10 mM KCl,5 mM ATP, 200 μM coenzyme A, 10 mM dithiothreitol and malatedehydrogenase. An aqueous solution of inhibitor was added (forinhibitors which were insoluble in water, a stock solution was preparedin DMSO. However the final DMSO concentration in the cuvette was notallowed to exceed 1%.). Finally, tripotassium citrate was added to 100μM final. This is K_(m) for citrate (Wells et al (Eur. J. Biochem.,1992, 204, 249-255) and Houston et al (Biochim. Biophys. Acta, 1985,844, 233-239)). Data analysis was performed using the curve fittingpackage Enzfitter (Elsevier Biosoft). For competitive inhibitors datawas fitted to the equation

    v=v.sub.max./(2+I/K.sub.i)

where v is the observed rate and I is the concentration of inhibitoradded. Thus the dissociation constant K_(i) for the inhibitor could befound.

Results:

The compound of Example 1 had a Ki of 0.8 μM (rat); and the compound ofExample 2 (lactone of Example 1) was inactive (rat).

The compounds of Examples 3 to 9 had Ki values (rat) of less than 30 μM.

The compounds of Examples 13 to 16 had Ki values (human) of less than 82μM.

The compounds of Example 18 had a Ki value of 0.76 μM (human) and 0.70μM (rat); and the compound of Example 20, a Ki value of 0.69 μM (human)and 0.70 μM (rat).

2. Measurement of effect of compounds on cholesterol (CL) and fatty acid(FA) synthesis in HepG2 Cells HepG2 cells were cultured in 24-well cellculture plates in DMEM (Dulbecco's Modified Eagle's Medium) containingHepes (20 mM), bicarbonate (10 mM), glutamine (2 mM) and foetal calfserum (10% w/v). Once the cells had grown to between 80% and 90%confluence, the medium was replaced by DMEM without the addition offoetal calf serum and the cells incubated overnight. The rates ofcholesterol and fatty acid synthesis were then measured by the additionof ³ H₂ O, to a specific radioactivity of 71 mCi/mmol, for the final 90min of the incubation. Vehicle or test compound were added to the mediumeither 1.5 or 14.5 hr prior to the addition of ³ H₂ O to give the finaldesired concentration. Incubations were terminated and the rates ofcholesterol and fatty acid synthesis determined from the amounts of ³ Hincorporated into cellular cholesterol and fatty acids as describedpreviously (Berkhout et al.; Biochem J., 1990, 272, 181).

Results

    ______________________________________                                                     Conc. CL synthesis                                                                             FA synthesis                                                 (μM)                                                                             % of control                                                                             % of control                                    ______________________________________                                        Compound of Example 1                                                                        1000    80 ± 6  123 ± 3                                  Compound of Example 2                                                                        30      8 ± 3   32 ± 3                                   (lactone of Example 1)                                                        Compound of Example 11                                                                        3      69 ± 13 74 ± 11                                  Compound of Example 11                                                                       10      86 ± 29 76 ± 13                                  ______________________________________                                    

3. Measurement of hyperlipidaemic activity in rats and dogs

(a) Spmgue Dawley Rat

The compound of example 2 was administered to Spmgue Dawley rats intheir diet at a concentration of 0.125% (w:w) for 7 days. Measurement ofplasma cholesterol levels and triglyceride levels by standard techniquesindicated that the compound had reduced plasma cholesterol levels by 30%and plasma triglyceride levels by 64%.

25 (b) Dog The compound of example 2 was administered to male Beagledogs at a level of 25 mg/kg/day for 2 weeks. Measurement of plasmacholesterol levels and triglyceride levels by standard techniquesindicated that the compound had reduced plasma cholesterol levels by20-25% and plasma triglyceride levels by 20-25%.

We claim:
 1. A compound of structure (I): ##STR25## in which, each groupR¹ is independently a lipophilic and/or electron withdrawing group; n is5 to 8; andeither R² and R³ are both hydrogen, R⁴ is hydrogen or hydroxyand R⁵ is CH(R⁶)R⁷ in which R⁶ is hydrogen or hydroxy and R⁷ is acarboxyl group or a carboxylic acid ester group hydrolysable to acarboxyl group; or R⁴ is hydrogen and R⁵ is hydrogen or hydroxy, R² ishydroxy and R³ is a carboxyl group or a carboxylic acid ester grouphydrolysable to a carboxyl group; or R² and R³ are hydrogen and R⁴ andR⁵ together form a group ═C(R⁶)R⁷ in which R⁶ and R⁷ are as definedabove, or a salt thereof.
 2. A compound according to claim 1 in which R²and R³ are both hydrogen, R⁴ is hydroxy and R⁵ is CH(R⁶)CO₂ H in whichR⁶ is hydrogen.
 3. A compound according to claim 1 in which R⁴ ishydrogen, R⁵ is hydrogen or hydroxy, R² is hydroxy and R³ is CO₂ H.
 4. Acompound of structure (IB): ##STR26## in which each group R^(1b) isindependently a lipophilic and/or electron withdrawing group; n^(b) is 5to 8; andeither R^(2b) and R^(3b) are both hydrogen, R^(4b) is hydrogenor hydroxy and R^(5b) is CH(R^(6b))CO₂ H in which R^(6b) is hydrogen orhydroxy; or R^(4b) is hydrogen and R^(5b) is hydrogen or hydroxy, R^(2b)is hydroxy and R^(3b) is CO₂ H; or R^(2b) and R^(3b) are hydrogen andR^(4b) and R^(5b) together form a group ═C(R^(6b))CO₂ H, or apharmaceutically acceptable salt thereof.
 5. A compound according toclaim 4 in which R^(2b) and R^(3b) are both hydrogen, R^(4b) is hydroxy,R^(5b) is (CHR^(6b))CO₂ H in which R^(6b) is hydrogen.
 6. A compound ofclaim 1 thatis:±(3R*,5S*)3-carboxymethyl-5-[6-(2,4-dichlorophenyl)hexyl]-3-hydroxytetrahydrofuran-2-one,±(4R*,5S*)4-carboxy-5-[8-(2,4-dichlorophenyl)octyl]-4-hydroxytetrahydrofuran-2-one,±(4R*,5R*)4-carboxy-5-[8-(2,4-dichlorophenyl)octyl]-4-hydroxytetrahydrofuran-2-one,±(3R*,5R*)3-(carboxymethyl)-5-[6-(2,4-dichlorophenyl)-hexyl]tetrahydrofuran-2-one,(+)(3R*,5S*)3-carboxymethyl-5-[6-(2,4-dichlorophenyl)-hexyl]-3-hydroxytetrahydrofuran-2-oneor(-)(3R*,5S*)3-carboxymethyl-5-[6-(2,4-dichlorophenyl)-hexyl]-3-hydroxytetrahydrofuran-2-one.7. A compound of claim 4 thatis:±(3R*,5S*)3-carboxy-11-(2,4-dichlorophenyl)-3,5-dihydroxyundecanoicacid, disodium salt,(+)(3R*,5S*)3-carboxy-11-(2,4-dichlorophenyl)-3,5-dihydroxyundecanoicacid, disodium salt,(+)(3R*,5S*)3-carboxy-11-(2,4-dichlorophenyl)-3,5-dihydroxyundecanoicacid, disodium salt,±(E)-3-carboxy-11-(2,4-dichlorophenyl)-5-hydroxy-2-undecenoic acid,±(2R*,3R*,5S*)3-carboxy-11-(2,4-dichlorophenyl)-2,3,5-trihydroxyundecanoicacid, disodium salt,±(2R*,3S*,5R*)3-carboxy-11-(2,4-dichlorophenyl)-2,3,5-trihydroxyundecanoicacid, disodium salt, or±(3R*,5S*)3-carboxy-11-(4-chloro-2-trifluoromethylphenyl)-3,5-dihydroxyundecanoicacid, disodium salt.
 8. A pharmaceutical composition comprising acompound of structure (I) as defined in claim 1, in association with apharmaceutically acceptable carrier.
 9. A method of lowering serumtriglyceride and cholesterol levels which comprises administering to amammal in need thereof an effective amount of a compound of structure(I) as defined in claim
 1. 10. A method of lowering serum triglycerideand cholesterol levels which comprises administering to a mammal in needthereof an effective amount of a compound of structure (IB) as definedin claim 4, or a pharmaceutically acceptable salt thereof.